Faecal transplantation and Clostridioides difficile infection

Faecal microbiota transplantation (FMT), known equally well as faecal transplantation or faecal bacteriotherapy, is the process of implanting the faecal suspension containing balanced microbiota from a healthy donor to the colon of a recipient patient. Excessive growth of Clostridioides difficile (C difficile) in the intestinal microbiota resulting from antibiotic consumption is currently a rising threat to public health. FMT is one of the most important, newer approaches to treating C difficile infections. Since C difficile is regarded as an opportunistic bacterium triggering disease in conditions of disturbed homeostasis of the intestinal microbiota, restoration of healthy intestinal microflora facilitates suppression of toxic strain of C difficile by anaerobic bacteria of normal intestinal microflora with concomitant cure. Nurses have important role in caring for patients after faecal transplantation.

Особливості отримання щільної фекальної культури яєць гельмінтів роду Trichuris, виділених від овець

Наведено результати досліджень щодо ефективності запропонованого способу отримання щільної фекальної культури яєць нематод роду Trichuris, виділених від овець. Ефективність запропонованого способу за різних режимів відстоювання фекальної суспензії (10, 15 та 20 хв.) відносно показника кількості отриманих яєць нематод перевищує флотаційно-центрифужний та загальновідомий методи на 30,44–56,55 % та 12,65–45,44 % відповідно. Удосконалений спосіб дає змогу скоротити витрати часу на проведення методики порівняно із флотаційно-центрифужним (на 8,28–25,03 %) та загальновідомим методами (на 4,70–16,77 %). The results of research on the effectiveness of the proposed method for obtaining a dense faecal of nematodes’ eggs of Trichuris genus in sheep are presented. The effectiveness of our method for different modes of settling the faecal suspension (10, 15 and 20 min) concerning to the number of nematode eggs obtained exceeds the flotation-centrifugal and well-known methods by 30.44–56.55% and 12.65–45.44% respectively. The improved method allows to reduce time loss on conducting the procedure compared with flotation-centrifuge (by 8.28–25.03%) and well-known methods (4.70–16.77%). For the purpose of successful fight against parasitosis, scientists of the world annually develop and produce a large number of drugs and formulations of chemical and biological origin that are pre-tested with a lot of approbation. One of such approbation is the use of experimental parasitosis simulation in the conditions of in vitro. Egg nature of causative agents of parasitic diseases is often used in the development of new methods for lifelong diagnosis and conducting comparative effectiveness of existing ones. However, in the literature, insufficient information has been found on the methods for obtaining eggs of helminths. In this regard, the aim of the research is to improve the method of obtaining a dense fecal eggs of worms of the Trichuris genus in sheep. The research task is to determine the effectiveness of the proposed method in comparison with the well-known ones. The research has been conducted on the base of the Laboratory of Parasitology and Veterinary-Sanitary Expertise at Poltava State Agrarian Academy. The eggs of trichuroses are obtained in three ways: a flotation-centrifuge method for cultivating eggs of Trichuris muris; a well-known method of cultivating eggs Trichuris muris; the proposed method with different terms for fecal suspension (10, 15 and 20 min). By testing a flotation-centrifuge method for obtaining trichurise eggs from sheep, it has been found that the average number of washed eggs is 15.72±0.67. Using a well-known method, it was found that the average obtained number of eggs is 19.74±0.74 specimens. The use of our method for different modes of settling the fecal suspension (10, 15 and 20 min) resulted in an average of 22.66±0.59, 36.18±1.50 and 28.10±0.90 eggs. Conclusions: The improved method has a higher efficiency in terms of the number of eggs obtained compared to flotation-centrifuge (30.44 to 56.55%) and well-known (12.65 to 45.44%) methods. It has been found out that the efficiency of the offered method exceeds the efficiency of the flotation-centrifugal (by 8.28–25.03%) and the well-known (on 4.70–16.77%) methods in terms of time loss.

Electrical cream separator coupled with vacuum filtration for the purification of eimerian oocysts and trichostrongylid eggs

Several methods have been proposed for separation of eimerian oocysts and trichostrongylid eggs from extraneous debris; however, these methods have been considered to be still inconvenient in terms of time and wide-ranging applications. We describe herein an alternative way using the combination of electrical cream separator and vacuum filtration for harvesting and purifying eimerian oocysts and haemonchine eggs on large-scale applications with approximately 81% and 92% recovery rates for oocysts and nematode eggs obtained from avian and ovine faeces, correspondingly. The sporulation percentages as a measure of viability in the harvested oocysts and eggs from dry faecal materials are nearly 68% and 74%, respectively, and 12 liters of faecal suspension can be processed in approximately 7.5 min. The mode of separation in terms of costs (i.e. simple laboratory equipments and comparably cheap reagents) and benefits renders the reported procedure an appropriate pursuit to harvest and purify parasite oocysts and eggs on a large scale in the shortest duration from diverse volumes of environmental samples compared to the modified traditional sucrose gradient, which can be employed on a small scale.

G and P types of bovine group A rotavirus in northern India

Animal rotaviruses (RVs) are considered as a potential threat to humans due to possibility of interspecies transmission and exchange of genomic materials. Among several RVs, the group A rotaviruses (RVA) are the major cause of diarrhoea in cattle and buffalo calves worldwide. The present study was carried out to understand epidemiology and types of RVA circulating in Mathura region of northern India. One hundred faecal samples were collected from diarrhoeic cattle (n=94) and buffalo calves (n=6) from organized dairy farms. Viral RNA was extracted from faecal suspension and was transcribed to cDNA using RT-PCR. The cDNA was amplified for VP6, VP7 and VP4 genes and multiplex nested PCR was done for G and P genotyping. Twelve samples were detected positive for RVA by antigen detection ELISA and eleven samples produced expected amplicon for group A specific VP6 gene. On genotyping with G3, G6, G8 and G10 specific primers for VP7 gene and P [1] and P [11] specific primers for VP4 gene, G6 genotype predominated over other genotypes. Dual genotypes were also observed. In P typing only one isolate was found to be positive for P [1] and rest all samples remained untypable. The result of present study indicates G6 as a major G genotype and change in frequencies of distribution of G types in this part of the country.

Relative efficacy of inocula from rumen fluid or faecal solution for determining in vitro digestibility and gas production

Three experiments were conducted to study the effects of substituting rumen fluid (RF) with faecal suspension (FS) as an inoculum for determination of in vitro dry matter digestibility (IVDMD, experiment 1) and gas production (exp. 2). Barley grain (BG), Persian clover (PC), alfalfa (ALF), bromegrass (BR) and barley straw (BS) were used to evaluate the efficacy of the two inocula. In exp. 1, IVDMD was determined using RF or FS containing 3, 6 and 9% fresh cattle faeces as inocula. Except for BS, IVDMD values obtained with either RF or FS containing 3% faeces were not different (P > 0.05). In contrast, FS containing 6 or 9% faeces resulted in lower (P < 0.05) IVDMD than those obtained with RF. Total VFA, butyric and valeric acids of in vitro supernatant were higher (P < 0.05) in RF than FS incubated buffers. Molar proportions of acetic and propionic acids were variable among feeds. Mean acetic:propionic ratio was similar (except for BG) for RF or FS. In exp. 2, substitution of RF with FS containing 6, 11 or 16% cattle faeces as inoculum generally resulted in an increased lag time for four feeds (PC was not included). The rate of gas production was lower when FS was used instead of RF for ALF and BR. For BG however, use of FS resulted in a higher (P < 0.05) rate of gas production than when RF was used. Total gas production values were similar within all the feeds regardless of the source of inoculum. In experiment 3, the relative size of the total and cellulolytic populations were higher for RF than for FS, with a greater diversity of genera of bacteria isolated from RF. Results from this study indicate that FS has the potential to be used instead of RF to obtain IVDMD and gas production for BG and forage hay but not for poor quality roughages such as BS. However, more research with FS is required to confirm these findings. Key words: IVDMD, gas production, rumen fluid, faecal suspension

Experimental observations on the pathogenesis of necrobacillosis

SUMMARYEarlier studies showed that the minimum infective dose (>106 organisms) of a virulent strain of Fusobacterium necrophorum could be greatly reduced by suspending the fusobacteria in sub-lethal doses of cultures of other bacteria such as Escherichia coli before inoculating mice subcutaneously.In the present study the infective dose of the same strain of F. necrophorum was reduced by a factor of >103 by suspending the fusobacteria in sub-lethal doses of 5% homogenate of gaur or wallaby faeces. Sterile faecal filtrate had no such effect. The sites of low grade infection produced by the prior subcutaneous injection of E. coli culture or gaur faecal suspension were susceptible to superinfection by doses of F. necrophorum far below those required to infect normal tissue.This work helps to explain the production of necrobacillosis by the faecal contamination of small wounds. It proved impossible, however, to produce necrobacillosis in mice by the subcutaneous injection of faecal suspensions from 33 farm cattle. This suggests that the proportion of cattle with virulent F. necrophorum in their faeces is low.

Effects of antibiotics on the survival ofSalmonellain the American cockroach

SUMMARYThe faecal survival ofSalmonella typhimuriumorally administered to restrained American cockroaches maintained on an antibiotic diet was investigated. Significant reductions in median numbers of total aerobic faecal microorganisms were noted in insects fed antibiotics daily, and whenSalmonellawas introduced to this modified micro-environment, it persisted for 44 days in all specimens. Multiplication of the pathogen was also observed in these insects, with numbers often exceeding 103times that of the initial input. This differs significantly from our previous results showing that the pathogen is unable to multiply in conventional cockroaches. Attempts to restore the normal flora by feeding a faecal suspension from untreated cockroaches resulted in a decrease in numbers ofSalmonellaexcreted, but did not result in their elimination. Carcasses of infected cockroaches retained viableSalmonellafor at least 60 dayspost mortem, or 104 days after the infective meal.